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Enhanced Calvarial Bone Repair Using ASCs Engineered with RNA-Guided Split dCas12a System that Co-Activates Sox 5, Sox6, and Long Non-Coding RNA H19. | LitMetric

AI Article Synopsis

  • A new method using an RNA-guided Split dCas12a system has been developed to activate long non-coding RNA H19 in stem cells to promote bone healing, especially for large cranial defects.
  • Previous attempts to activate H19 alone in adipose-derived stem cells (ASCs) did not result in effective chondrogenesis, leading researchers to combine it with other key transcription factors, Sox5 and Sox6.
  • Co-activating H19 along with Sox5 and Sox6 in ASCs resulted in significantly better cartilage formation and bone repair, demonstrating the potential of this system for improving tissue regeneration.

Article Abstract

Healing of large calvarial bone defects remains challenging. An RNA-guided Split dCas12a system is previously harnessed to activate long non-coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow-derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose-derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co-activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co-activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co-activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration.

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Source
http://dx.doi.org/10.1002/smll.202306612DOI Listing

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